In order to definitude the influence caused by the different sampling in voice assessment.Method:We comparing the results acquired by total section and subsection sampling.Result:The results acquired by subsection tended to normal more than those acquired by total section. Conclusion:Subsection sampling voice assessment might conceal the drgree of the disease state of patients

Objective:This study aimed to investigate the possible mechanism of 32P colloid induced apoptosis of craniopharyngi-oma (CP) cells in vitro and the relationship between dose effect and time effect. Methods:This study established a primary cell culture of CP limited subculture cell line. Methyl thiazolyl tetrazolium (MTT) assay was performed to plot the cell survival curve after the CP cells were treated with 32P colloid at different concentrations and time. Apoptotic rate was detected by flow cytometry(FCM). Apoptosis related DNA was investigated by TUNEL fluorescent staining. The morphological characteristics of apoptotic cells were determined by Hoechst33342 fluorescence staining. The ultrastructure of apoptotic cells was investigated by transmission electron microscopy (TEM). Results:Hoechst33342 fluorescence staining, TUNEL fluorescence staining, and TEM revealed that 32P colloid induced the apoptosis of CP cells. 32P colloid reduced the survival rate and increased the apoptotic rate of CP cells as concentration (0 MBq/mL to 14.80 MBq/mL) and time (1 d to 14 d) were increased. Conclusion: 32P colloid could effectively inhibit the growth of CP cells and induce apoptosis in vitro. High concentrations and prolonged time could induce a remarkable effect.

Objective To establish human embryonic stem cells (hESCs) feeder-independent and cell factor-free culture system. Methods Effect of high and low clone densities of hESCs culture was compared and impact of the clone densities to hESCs culture media was analyzed. Results HESCs could maintain their undifferentiated states at high clone density (34 clones/cm2) without cell factors. At the same time,the bone morphology protein (BMP)-like induction of N2 and B27 supplements (NB) medium could be modulated by the clone density,and high level of BMP-like induction was accompanied by high clone density. Conclusion High clone density of hESCs can change the environments by themselves to maintain the undifferentiated states,which provides a new clue to explore the mechanism of undifferentiated states of hESCs and simplify the culture medium.

Objective To explore the mechanism of LMWH therapy for SAP.Methods 48 wistar rats were random divided into 3 groups,sham group(S group),severe acute pancreatitis group(SAP group)and LMWH therapy group(H group).Serum amylase,IL-6,acinar cell apoptosis and the activity of NF-κB were detected and compared.Results The expression of amylase and IL-6 in SAP group was significantly higher than that in H group(P＜0.01).The apoptosis index of acinar cell in SAP group wag significantly lower than that in H group(P＜0.01),while the activity of NF-κB in SAP groupwas stronger than that in H group.Conclusions LMWH therapy may ameliorate SAP by inducing acinar cell apoptosis through suppressing the activity of NF-κB.

Objective To investigate the diagnostic value of magnetic resonance imaging(MRI) conventional sequences and diffusion weighted imaging (DWI) in polycystic ovary syndrome(PCOS).Methods 75 patients with clinical-confirmed PCOS and 46 healthy women in Yuyao Traditional Chinese Medicine Hospital from Oct.2016 to Apr.2018 were selected.All the subjects received MRI conventional sequences and DWI examination.The size of ovary and number of follicles were observed,and apparent diffusion coefficient (ADC) values in ovarian stroma were measured.The ovarian morphologic changes and ADC values between the two groups were compared.Results The volume of ovary and number of follicles in the control group were (6.6±1.5)cm3 and 6.9±1.8,the difference was statistically significant compared with that in PCOS group [(11.8±2.8)cm3,13.6±3.6)](P＜0.05).ADC value in ovarian stroma in PCOS group was reduced,and the difference was also statistically significant compared with that in the control group(P＜0.05).The sensitivity and specificity of diagnosing PCOS with ADC threshold of 1.39×10-3 mm2/s were 82.2％ and 68.9％,respectively.Conclusion MRI conventional sequences and DWI are helpful in diagnosis of PCOS,which has important significance for the clinical treatment and prognosis evaluation.

Objective:To analyse the risk factors of biliary leakage after surgical resection in patients with perihilar cholangiocarcinoma (PHCC).Methods:The medical data on 179 patients who underwent surgical resection for PHCC at the Department of Hepatopancreatobiliary Surgery, Ningbo Medical Centre Lihuili Hospital, Ningbo University from April 2000 to April 2020 were collected, and 160 patients were finally enrolled into this study. There were 86 males and 74 females, aged (63.4±10.8) years. The 44 patients with class B biliary leakage and the 5 patients with class C biliary leakage were classified into the biliary leakage group, while the remaining 111 patients were classified into the control group. Risk factors of biliary leakage were analysed by univariate and multivariate logistic regression analyses.Results:Operation time ≥360 min, resection and reconstruction of hepatic hilar vessels on the preserved side of liver and number of bile duct openings of >3 in remnant liver were significantly higher in the biliary leakage than the control group (all P<0.05). Multivariate analysis showed that resection and reconstruction of hepatic hilar vessels on the preserved side ( OR=2.322, 95% CI: 1.078-5.002, P=0.028) and 3 or more bile duct openings in the remnant liver ( OR=2.656, 95% CI: 1.198-5.892, P=0.016) were significantly associated with biliary leakage. Conclusion:Resection and reconstruction of hepatic hilar vessels on the preserved side of liver and 3 or more bile duct openings in remnant liver were independent risk factors for biliary leakage after PHCC resection.

Objective:To evaluate clinical and pathological factors related to the actual 5-year survival of patients with hilar cholangiocarcinoma (HCC).Methods:A total of 94 HCC patients who underwent radical surgery at the Department of Hepatobiliary and Pancreatic Surgery, Li Huili Hospital of Ningbo Medical Center from Jan 2000 to Jun 2015 were enrolled in this study.Patients were divided into two groups: postoperative survival group beyond 5 years and death group within 5 years. The clinical and pathological features of the two groups were analyzed.Results:Of the 94 patients, 19 (20.2%) had a postoperative survival time of more than 5 years. The actual 5-year overall survival rate of HCC patients (20.2%) was lower than that estimated by Kaplan-Meier survival analysis (22.2%). Gender, age, CEA value, CA199 value, total bilirubin, Child-Pugh classification, Bismuth classification and preoperative jaundice reduction were not significantly different between the two groups nor there were significant difference between two groups in operation time, blood loss, surgical procedure, combined caudate lobectomy, combined pancreaticoduodenectomy, combined resection of surrounding organs, vascular reconstruction and number of bile duct orifices in remnant liver surface. There were significant differences between two groups in the variables of pathological phenotype ( P=0.012), lymph node metastasis ( P=0.001) and resection level ( P=0.048). Conclusion:Non-papillary type, lymph node metastasis and R 1 resection are the independent risk factors of the actual 5-year survival.

Nanomaterial-based drug sustainable release systems have been tentatively applied to bone regeneration. They, however, still face disadvantages of high toxicity, low biocompatibility, and low drug-load capacity. In view of the low toxicity and high biocompatibility of polymer nanomaterials and the excellent load capacity of hollow nanomaterials with high specific surface area, we evaluated the hollow polydopamine nanoparticles (HPDA NPs), in order to find an optimal system to effectively deliver the osteogenic drugs to improve treatment of bone defect. Data demonstrated that the HPDA NPs synthesized herein could efficiently load four types of osteogenic drugs and the drugs can effectively release from the HPDA NPs for a relatively longer time in vitro and in vivo with low toxicity and high biocompatibility. Results of qRT-PCR, ALP, and alizarin red S staining showed that drugs released from the HPDA NPs could promote osteogenic differentiation and proliferation of rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. Image data from micro-CT and H&E staining showed that all four osteogenic drugs released from the HPDA NPs effectively promoted bone regeneration in the defect of tooth extraction fossa in vivo, especially tacrolimus. These results suggest that the HPDA NPs, the biodegradable hollow polymer nanoparticles with high drug load rate and sustainable release ability, have good prospect to treat the bone defect in future clinical practice.
Animals ; Bone Regeneration ; Indoles ; Nanoparticles ; Osteogenesis ; Pharmaceutical Preparations ; Polymers ; Rats

Ulcerative Colitis (UC) has been reported to be related to Porphyromonas gingivalis (P. gingivalis). Porphyromonas gingivalis peptidylarginine deiminase (PPAD), a virulence factor released by P. gingivalis, is known to induce inflammatory responses. To explore the pathological relationships between PPAD and UC, we used homologous recombination technology to construct a P. gingivalis strain in which the PPAD gene was deleted (Δppad) and a Δppad strain in which the PPAD gene was restored (comΔppad). C57BL/6 mice were orally gavaged with saline, P. gingivalis, Δppad, or comΔppad twice a week for the entire 40 days (days 0-40), and then, UC was induced by dextran sodium sulfate (DSS) solution for 10 days (days 31-40). P. gingivalis and comΔppad exacerbated DDS-induced colitis, which was determined by assessing the parameters of colon length, disease activity index, and histological activity index, but Δppad failed to exacerbate DDS-induced colitis. Flow cytometry and ELISA revealed that compared with Δppad, P. gingivalis, and comΔppad increased T helper 17 (Th17) cell numbers and interleukin (IL)-17 production but decreased regulatory T cells (Tregs) numbers and IL-10 production in the spleens of mice with UC. We also cocultured P. gingivalis, Δppad, or comΔppad with T lymphocytes in vitro and found that P. gingivalis and comΔppad significantly increased Th17 cell numbers and decreased Treg cell numbers. Immunofluorescence staining of colon tissue paraffin sections also confirmed these results. The results suggested that P. gingivalis exacerbated the severity of UC in part via PPAD.
Animals ; Colitis, Ulcerative/microbiology* ; Mice ; Mice, Inbred C57BL ; Porphyromonas gingivalis/pathogenicity* ; Protein-Arginine Deiminases ; Virulence Factors

Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.